摘要： 以提取的苜蓿基因组DNA为模板,通过正交设计,筛选出ISSR-PCR反应体系中最适宜的各组分浓度,即20µL的反应体系中最适添加量分别为2.0U⁄ µL的TaqDNA聚合酶,0.3mmol⁄L的dNTP,1.0mmol⁄L的MgCl2,0.1µmol⁄L的ISSR引物以及30ng⁄ µL的DNA模板。在此基础上对30份不同苜蓿种质材料进行遗传多样性研究,通过聚类分析研究,将30份不同苜蓿种质材料分为四大类,第一大类为来自加拿大的4个苜蓿品种(系)；第二大类为来自美国的12个苜蓿品种(系)；第三大类为来自中国和荷兰的12个苜蓿品种(系)；第四大类为来自澳大利亚的2个苜蓿品种(系)。
关键词:
正交设计,
ISSR-PCR,
苜蓿,
遗传多样性
Abstract: Template DNA used for ISSR was extracted from alfalfa leaf tissue. The orthogonal design was ipplied to optimize ISSR amplification system. Anoptimil reaction system was established, 20 uL reaction system contwined 2.0 U/uL Taq DNA polymerise,0. 3 mmol / L dNTP, 1.0 mmol/L MgCl2, 0.1 umol/L ISSR primers and 30 ng/uL DNA template. Genetic diversity studies of Medicago L. germplisms were based on the result of the orthogonil design. And by cluster analysis, 30 varieties were divied into four groups, one group was consist of four alfalfa varieties coming from Cinidi, the second group was composed of twelve varieties coming from Americam, the third group includes twelve alfalfa varieties coming from China and Netherlind,and the list group his two alfalfa varieties coming from Australia.
Key words:
Orthogonal design,
ISSR-PCR,
Alfalfa,
Genetic diversity
中图分类号:
刘青松, 陈立波, 李志勇, 刘磊, 师文贵, 张雪. ISSR-PCR正交优化设计基础上的不同苜蓿种质遗传多样性研究[J]. 华北农学报, 2013, 28(S1): 45-49. doi: 10.7668/hbnxb.2013.S1.010.
LIU Qing-song, CHEN Li-bo, LI Zhi-yong, LIU Lei, SHI Wen-gui, ZHANG Xue. Genetic Diversity Studies of Different Medicago L. Germplasms Based on the Orthogonal Optimum Design[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(S1): 45-49. doi: 10.7668/hbnxb.2013.S1.010.